Genomic Nucleosome Organization Reconstituted with Pure Proteins
Research output: Contribution to journal › Journal article › Research › peer-review
Chromatin remodelers regulate genes by organizing nucleosomes around promoters, but their individual contributions are obfuscated by the complex in vivo milieu of factor redundancy and indirect effects. Genome-wide reconstitution of promoter nucleosome organization with purified proteins resolves this problem and is therefore a critical goal. Here, we reconstitute four stages of nucleosome architecture using purified components: yeast genomic DNA, histones, sequence-specific Abf1/Reb1, and remodelers RSC, ISW2, INO80, and ISW1a. We identify direct, specific, and sufficient contributions that in vivo observations validate. First, RSC clears promoters by translating poly(dA:dT) into directional nucleosome removal. Second, partial redundancy is recapitulated where INO80 alone, or ISW2 at Abf1/Reb1sites, positions +1 nucleosomes. Third, INO80 and ISW2 each align downstream nucleosomal arrays. Fourth, ISW1a tightens the spacing to canonical repeat lengths. Such a minimal set of rules and proteins establishes core mechanisms by which promoter chromatin architecture arises through a blend of redundancy and specialization.
Original language | English |
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Journal | Cell |
Volume | 167 |
Issue number | 3 |
Pages (from-to) | 709-721.e12 |
ISSN | 0092-8674 |
DOIs | |
Publication status | Published - 20 Oct 2016 |
Externally published | Yes |
Bibliographical note
Copyright © 2016 Elsevier Inc. All rights reserved.
- Adenosine Triphosphatases/chemistry, Chromatin/chemistry, Chromatin Assembly and Disassembly, DNA, Fungal/chemistry, DNA-Binding Proteins/chemistry, Genome, Fungal, Histones/chemistry, Nucleosomes/chemistry, Poly dA-dT/chemistry, Protein Biosynthesis, Saccharomyces cerevisiae/genetics, Saccharomyces cerevisiae Proteins/chemistry, Transcription Factors/chemistry
Research areas
ID: 301927082